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Abstract: SA-PO411

NLRP3 Inflammasome Activation Mediated by Lysyl Oxidase-Like 2 in Human Podocytes in Diabetic Condition

Session Information

Category: Diabetic Kidney Disease

  • 701 Diabetic Kidney Disease: Basic

Authors

  • Choi, Hoon Young, Yonsei University College of Medicine, Seodaemun-gu, Korea (the Republic of)
  • Jeon, Nara, Yonsei University College of Medicine, Seodaemun-gu, Korea (the Republic of)
  • Lim, Beom Jin, Yonsei University College of Medicine, Seodaemun-gu, Korea (the Republic of)
Background

Diabetic kidney disease (DKD), a major complication of diabetes mellitus is a most common cause of kidney failure. Sterile inflammation is a hallmark of DKD. The NLR family pyrin domain containing 3 (NLRP3) inflammasome is a mechanism of sterile inflammation known to be activated by metabolic stimuli and reactive metabolites associated with DKD. Recently, lysyl oxidase-like 2 (LOXL2) has been reported to associate with tissue fibrosis, inflammation, and oxidative stress, all of which are implicated in the pathogenesis of diabetes and its complications. In this study, we investigated the expression of LOXL2 in human kidney and podocyte, and its contribution to NLRP3 inflammasome expression in podocyte with high-glucose condition.

Methods

We evaluated the expression of LOXL2 in human kidney using immunoflurorescence staining. Real-time PCR and western blotting analysis for LOXL2 mRNA and protein expression were performed using cultured human immortalized podocytes. After fully differentiated, cultured human podocytes were exposed to high glucose (HG) for 48 hours. Podocyte-specific LOXL2 knock-down cells using CRISPR-Cas9 was generated.

Results

By real-time PCR and immunofluorescence staining, LOXL2 expression was identified in human glomerulus and was significantly increased in that with diabetic kidney disease compared with normal control. mRNA of NLRP3 inflammasome expression was also significantly increased in podocyte with HG condition (6.79±0.32 vs. 1.60±0.26, P<0.05). Gene silencing of LOXL2 significantly reduced mRNA and protein expression of NLRP3 inflammasome in immortalized human podocytes with HG compared to controls. Western blot analysis showed that collagen I and phosphorylated Smad2 protein expression were significantly decreased in LOXL2 knock-down podocytes.

Conclusion

Our results showed that NLRP3 inflammasome activation may attenuated by gene silencing of LOXL2 in podocytes in diabetic condition.